Dressing and preparation method thereof
A scaffold material and solution technology, applied in the field of medical materials, can solve the problems of autologous stem cells, low proliferation, differentiation, adhesion and angiogenesis, slow wound healing, complicated operation, etc., so as to promote rapid healing, reduce immune rejection, Simple operation effect
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Embodiment 1
[0038] Example 1. Isolation of human umbilical cord mesenchymal stem cells
[0039] Take a piece of human umbilical cord tissue, spread it into a petri dish, add complete medium, and transfer to 5% CO 2 , 37°C, saturated humidity of 95% CO 2 cultured in an incubator. In the first 5-7 days of static culture, the color of the culture medium changes to light yellow, and the first rehydration is carried out. When at least 5 cells crawling out of each plate were observed, the medium and tissue pieces in the plate were sucked off with a pipette, and the medium was changed to obtain primary human umbilical cord mesenchymal stem cells (P0).
[0040] After 10-12 days of primary culture, some cells crawl out to form clones, continue to culture until at least 5 clones appear in the whole dish, digest the cells for passage. Observation of passaged cells under an inverted microscope, P3 generation human umbilical cord mesenchymal stem cells, the results are as follows figure 1 .
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Embodiment 2
[0042] Embodiment 2, identification of human umbilical cord mesenchymal stem cells
[0043] Take human umbilical cord mesenchymal stem cells in the logarithmic growth phase P2, suck out the medium, add 0.25% trypsin + 0.02% EDTA digestion solution for digestion, then stop the digestion with an appropriate amount of serum, and gently pipette into a single cell suspension . Centrifuge at 1000rpm for 10min, discard the supernatant, wash the cells 3 times with 4°C pre-cooled PBS, resuspend evenly, and adjust the cell concentration to about 10 5 pcs / mL-10 6 individual / mL.
[0044] Take 8 loading tubes, add 500 μL of single cell suspension to each tube, mark tube 1 as standard control, add 5 μL of PE-labeled mouse IgG 1 Antibody and 5 μL FITC-labeled mouse IgG 1 Antibody; add 5 μL FITC or PE-labeled mouse anti-human cell surface molecules CD59, CD105, CD45, CD90 antibody working solution to each other tube. At room temperature, protected from light, incubate for 20 minutes. Wa...
Embodiment 3
[0046] Embodiment 3, dressing
[0047] The dressing is composed of a cell-scaffold mixture and a growth factor solution, wherein the volume ratio of the cell-scaffold mixture to the growth factor solution is 10:1.
[0048] The cell-scaffold mixture is composed of scaffold material solution and human umbilical cord mesenchymal stem cell suspension, the volume ratio of the scaffold material solution and human umbilical cord mesenchymal stem cell suspension is 7:3; the human umbilical cord mesenchymal stem cell The cell concentration of human umbilical cord mesenchymal stem cells in the suspension was 1×10 6 each / mL; the scaffold material solution is composed of DMEM / F12 medium, type I mouse collagen, chitin, 6-chondroitin sulfate, trehalose and glutaraldehyde, and the type I mouse collagen accounts for 1.0v of the scaffold material solution / v%, chitin accounts for 2v / v% of the scaffold material solution, 6-chondroitin sulfate accounts for 5v / v% of the scaffold material solutio...
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